CAJ | 학술논문

建立了利用Waters2695高效液相色谱测定毕赤酵母发酵液中S-腺苷蛋氨酸（S-adenosylmethionine，SAM）含量的方法，对HPLC—UV法检测SAM含量的色谱条件进行优化，最佳色谱条件如下：All-tima C18柱，150mm×4．6mm，5μm，流动相为V（0．01mmoL／L甲酸铵）：V（甲醇）=97：3，流速0．6mL／min，柱温30℃，检测波长260nm．优化后的色谱分离条件可使样品中的SAM与杂质达到较佳分离，稳定性、精密度及重复性都较好，其RSD均小于1．5％．利用SAM标品制作标准曲线Y=32408606．0721X＋130181．2824，线性范围为0．012～1．2g／L，R2达到0．9998，平均回收率为100．027％～102．533％．此方法可有效地检测发醇液中S-腺苷蛋氨酸含量．
건립료이용Waters2695고효액상색보측정필적효모발효액중S-선감단안산（S-adenosylmethionine，SAM）함량적방법，대HPLC—UV법검측SAM함량적색보조건진행우화，최가색보조건여하：All-tima C18주，150mm×4．6mm，5μm，류동상위V（0．01mmoL／L갑산안）：V（갑순）=97：3，류속0．6mL／min，주온30℃，검측파장260nm．우화후적색보분리조건가사양품중적SAM여잡질체도교가분리，은정성、정밀도급중복성도교호，기RSD균소우1．5％．이용SAM표품제작표준곡선Y=32408606．0721X＋130181．2824，선성범위위0．012～1．2g／L，R2체도0．9998，평균회수솔위100．027％～102．533％．차방법가유효지검측발순액중S-선감단안산함량．
An HPLC determination method was developed for measuring S-adenosylmethionine (SAM) contents in culture broth of Pichia pastoris. The optimized HPLC conditions for SAM analysis were 0. 01 mmol/L ammonium formate solution ( pH = 3.5 ) - methylalcohol ( V: V = 97 : 3 ) as mobile phase, flow rate 0. 6 mL/min, column temperature 30℃, and detection wavelength 260 nm. The separation conditons described above achieved good separation effect, together with high stability, sensitivity, and reproducibility. The standard curve of SAM was Y = 32408606. 0721 X + 130181.2824 , and the linear range was 0. 012 × 1.2 g/L, R2 was 0. 9998, the average recycle rate was 100. 027% - 102. 533%. This method would be useful for SAM analysis during Pichia pastoris fermentation.